2,4-Dinitrophenylhydrazine Hydrochloride [for HPLC Labeling]

The compound 1 is an HPLC labeling reagent, which has a hydrazino group and easily reacts with a carbonyl group to form the corresponding hydrazones. The resultant hydrazone is stable enough to reach the detector without any decomposition under reversed phase HPLC. An excellent chromatogram can be obtained by measuring at 254 nm, an absorption wavelength that is generally used for UV detection.

Application examples:
[Aldehydes]
1 mg of a sample, 1 mg of the labeling reagent 1, 1 mL of methanol, and 0.5 mL of 1 NHCl are mixed. Close the cap of the reaction vessel and incubate the mixture at 40 °C for 10 min. After cooling to room temperature, use it as the HPLC sample solution.
[Keto acids] ^1,2)
A sample is dissolved in 1 mL of diluted HCl solution containing labeling reagent 1 (500 µmol / 2 N HCl 100 mL). Incubate the mixture at 30 °C for 30 min(The reactions are completed in 5 min and 20 min for ketomonocarboxylic acids and ketodicarboxylic acids,respectively.). It is preferable to add over 4 eq. amount of the labeling reagent, and resultant hydrazones can be extracted with ethyl acetate.
[Urine, 17-Ketosteroids in blood plasma] ^3,4)
A sample is dissolved into methanol, and acidified with 3~4 drops of conc. HCl. Excess amount of 0.2% labeling reagent 1 in methanol is added. Incubate the mixture at 50 °C for 5 min.
[Others]
Aliphatic carbonyl compounds^5,6）, aliphatic aldehydes^7~9)

TCI Products Pamphlet
HPLC Labeling Reagents ( PDF 10MB )